Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: rhIL-37 delays preterm delivery by reducing LPS-induced inflammatory effects and NLRP3 inflammasome activation in mice. ( A ) Orbital blood was collected from the control mice ( n = 6), LPS-induced preterm mice (LPS; n = 6), and rhIL-37 + LPS-co-treated preterm mice (rhIL-37 + LPS; n = 6).Serum levels of IL-1β, IL-6 and TNF-α were detected via ELISA. ( B ) Foetal membrane and placental tissues were collected from mice ( n = 6 per group) immediately before labour,, and the mRNA expression levels of NLRP3, Caspase-1 and ASC were detected via qRT-PCR. ( C ) Foetal membrane and placental tissues were collected from the mice in each group ( n = 3 per group), and the protein levels of NLRP3, Caspase-1, Pro-caspase-1 and ASC were analyzed by western blotting. ( D ) Western blot Results were normalised to β-actin. Bar graphs show statistical quantification of the western blot data. All data met the assumption of homogeneity of variance. Data are presented as mean ± s.e.m. Results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (one-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay, Membrane, Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: IL-37 knockdown enhances LPS-induced inflammatory and NLRP3 inflammasome activation in HTR-8/Svneo cells. ( A ) Cell supernatants were collected from control, si_NC- or si_IL37-treated cells following LPS exposure, and IL-37 expression level were detected by ELISA, qRT-PCR and Western blotting. ( B ) IL-1β, IL-6 and TNF-α levels in the cell supernatants were detected by ELISA. ( C ) The mRNA expression levels of NLRP3, Caspase-1 and ASC after cell treatment with LPS ( n = 5) were measured by qRT-PCR analysis. ( D ) NLRP3,pro-caspase-1, caspase-1 and ASC protein levels were detected by Western blotting, and the results were normalised to β-actin. All data exhibited equal variance.The data are shown as the mean ± s.e.m. The results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (One-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Knockdown, Activation Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: Nigericin reversed the anti-inflammatory effects and effects of rhIL-37 and its inhibition of NF-κB p65 activation in LPS-induced PTB.t ( A ) Cell supernatants were collected from HTR-8/Svneo cells co-treated with rhIL-37 and LPS, then either left untreated or treated with nigericin.The mRNA expression of NLRP3, caspase-1, and ASC was detected. ( B ) Cells were collected, and the protein expression of NLRP3, caspase-1, pro-caspase-1, and ASC was detected by western blotting. Results were normalised to β-actin and statistically analyzed.( C ) HTR-8/Svneo cell supernatants were collected after co-treatment with rhIL-37 and LPS (with or without nigericin), and the expression levels of IL-1β, IL-6, and TNF-α were detected by ELISA. ( D ) HTR-8/Svneo cells co-treated with rhIL-37 and LPS (with or without nigericin) were collected. The mRNA expression of p65 was detected by qRT-PCR, and protein expression of p65 and p-p65 was detected by western blotting. Densitometric quantification of p-p65 relative to p65 was performed. Statistical analysis of the western blot results was performed.All data exhibited equal variance.The data are shown as the means ± s.e.m. The results are representative of at least three independent experiments..

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Annals of Hematology

Article Title: LncRNA PVT1 targets miR-143-3p to modulate endothelial cell function and thereby participate in deep vein thrombosis (DVT) of the lower limbs

doi: 10.1007/s00277-026-06833-4

Figure Lengend Snippet: Effects of si-PVT1 transfection on CoCl 2 –induced angiogenesis and inflammatory levels in HUVECs. A. Changes in vascular endothelial growth factor (VEGF) expression in HUVECs following CoCl 2 induction and si-PVT1 transfection. B. Effects of CoCl 2 induction and LncRNA PVT1 inhibition on fibroblast growth factor-2 (FGF-2) expression in HUVECs. Effects of different transfection treatments on inflammatory cytokine expression: TNF-α (C) and IL-6 (D) . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: We used the following kits to assess the effects of different transfection treatments on endothelial cell angiogenesis and inflammatory levels: the Human VEGF ELISA Kit (97053ES, YENSEN, Shanghai), Human Fibroblast Growth Factor 2 (FGF2) ELISA Detection Kit (JL14546, JONLNBIO, Shanghai), Human TNF-α ELISA Kit (EK182, MULTI SCIENCES, Shanghai), and Human IL-6 ELISA Kit (EK106, MULTI SCIENCES, Shanghai).

Techniques: Transfection, Expressing, Inhibition

Journal: Annals of Hematology

Article Title: LncRNA PVT1 targets miR-143-3p to modulate endothelial cell function and thereby participate in deep vein thrombosis (DVT) of the lower limbs

doi: 10.1007/s00277-026-06833-4

Figure Lengend Snippet: Effects of miR-143-3p inhibitor on HUVEC function and inflammatory levels. Expression of LncRNA PVT1 (A) and miR-143-3p (B) in HUVECs following miR inhibitor transfection. C. Effect of miR inhibitor on viability of damaged HUVECs. D. Apoptotic changes in damaged HUVECs after miR inhibitor transfection. E. Effect of miR inhibitor on apoptosis-related gene expression in damaged HUVECs. F. Changes in VEGF and FG-2 expression in damaged HUVECs following miR inhibitor transfection. G. Effect of miR inhibitor on expression of inflammatory cytokines TNF-α and IL-6 in damaged HUVECs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: We used the following kits to assess the effects of different transfection treatments on endothelial cell angiogenesis and inflammatory levels: the Human VEGF ELISA Kit (97053ES, YENSEN, Shanghai), Human Fibroblast Growth Factor 2 (FGF2) ELISA Detection Kit (JL14546, JONLNBIO, Shanghai), Human TNF-α ELISA Kit (EK182, MULTI SCIENCES, Shanghai), and Human IL-6 ELISA Kit (EK106, MULTI SCIENCES, Shanghai).

Techniques: Expressing, Transfection, Gene Expression

Journal: Chinese Medicine

Article Title: Single-cell RNA sequencing reveals the therapeutic mechanism of Calvatia lilacina in promoting wound healing of anal fistula

doi: 10.1186/s13020-025-01293-w

Figure Lengend Snippet: The impact of CLS on inflammation, angiogenesis, and collagen at different time points. A – C ELISA detection of TNF-α, VEGF, and Collagen I expression levels in granulation tissue; D Immunohistochemical staining of CD14 and CD11b; E , F Immunohistochemical quantitative analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns: not significant ( P > 0.05) vs.the control group; n = 10

Article Snippet: TNF-α, COL1A1, VEGF Elisa Kit (EK182, EK183, EK1C01, Multi Sciences, China); WASF3 Antibody (67620-1-lg, Proteintech, China); α-SMA (19245s, CST, USA); CD14 Antibody (ab183322, abcam, Britain); Cluster of Differentiation 11b (CD11b) Antibody (ab133357, abcam, Britain); IL-6 flow Antibody (562050, BD Pharmingen, USA); JAK2 (WL02188, Wanleibio, China); p-JAK2 (WL02997, Wanleibio, China); STAT3 (WL03207, Wanleibio, China); p-STAT3 (WL06214, Wanleibio, China);

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemical staining, Staining, Control

Journal: Chinese Medicine

Article Title: Single-cell RNA sequencing reveals the therapeutic mechanism of Calvatia lilacina in promoting wound healing of anal fistula

doi: 10.1186/s13020-025-01293-w

Figure Lengend Snippet: Flow cytometry and SEM detection of adhesion and effect of CLS on macrophages. A Flow cytometry was used to determine the ratio of IL-6 positive cells in macrophage after extraction. B Quantitative analysis the rate of IL-6 positive macrophage. C , D Elisa detection of IL-6 and CXCL-8 in cell supernatant. E , F Intensity of intercellular communication between control group and treatment group. G Analysis of IL-6 + macrophage and fibroblast receptor-ligand binding. H Analysis of cell population interaction intensity of IL-6, CXCL-8 signaling pathway. * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: not significant ( P > 0.05) vs.the control group; n = 3

Article Snippet: TNF-α, COL1A1, VEGF Elisa Kit (EK182, EK183, EK1C01, Multi Sciences, China); WASF3 Antibody (67620-1-lg, Proteintech, China); α-SMA (19245s, CST, USA); CD14 Antibody (ab183322, abcam, Britain); Cluster of Differentiation 11b (CD11b) Antibody (ab133357, abcam, Britain); IL-6 flow Antibody (562050, BD Pharmingen, USA); JAK2 (WL02188, Wanleibio, China); p-JAK2 (WL02997, Wanleibio, China); STAT3 (WL03207, Wanleibio, China); p-STAT3 (WL06214, Wanleibio, China);

Techniques: Flow Cytometry, Extraction, Enzyme-linked Immunosorbent Assay, Control, Ligand Binding Assay

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A , B Western blot analysis of HIF-1α, BNIP3, NIX, MFN1, DRP1, LC3B, and p62 protein levels in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. C Autophagosomes were analyzed in M7/8+shCTR ( n = 2), M9a+shCTR ( n = 2), and M9a+sh EPAS1 ( n = 2) cells using TEM under hypoxia. Scale bar: 1 μm. D Autophagic flux was assessed by quantifying the colocalization of LC3B (green) and Mitotracker (red) (marking mitophagosomes) using immunofluorescence in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. E Cytoplasmic free mtDNA was determined by staining the double-stranded DNA (dsDNA) (green) and Mitotracker (red) using immunofluorescence in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. F Western blot analysis of NLRP3, IL1β, cGAS, STING, P65, and P-P65 protein levels in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. G Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under normoxic and hypoxic conditions using an ELISA kit. All cells were cultured under normoxia (21% O₂) or hypoxia (1% O₂) for 48 h.#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. One-way ANOVA followed by Tukey’s test; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Total ROS and mtROS contents were determined in the M9a+shCTR ( n = 4), M9a+sh EPAS1 ( n = 4) and M9a+sh EPAS1 -NAC ( n = 4). The mtROS and total ROS contents in the M9a+sh EPAS1 and M9a+sh EPAS1 -NAC cells were normalized to those in the M9a+shCTR cells. B Western blot analysis of HIF-1α, BNIP3, NIX, LC3B, and p62 protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. C Autophagic flux was quantified by colocalizing LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in the three cell lines under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP content were measured in the three cell lines under hypoxia. MMP and ATP levels in M9a+sh EPAS1 and M9a+sh EPAS1 -NAC cells were normalized to those in M9a+shCTR cells. G CCK-8 assay showing cell viability in the three cell lines under hypoxia. H Cell apoptosis was determined using Annexin V-FITC staining in the three cell lines under hypoxia. I Western blot analysis of C-Caspase-3 and BAX protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) using immunofluorescence in the three cell lines under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium supernatant were measured in the three cell lines under hypoxia using an ELISA kit. The three cell lines, M9a+shCTR ( n = 4), M9a+sh EPAS1 ( n = 4), and M9a+sh EPAS1 -NAC ( n = 4), were treated with hypoxia (1% O₂) for 48 h, with NAC treatment (10 mM) administered during the last 24 h.#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. One-way ANOVA followed by Dunnett’s test; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Schematic diagram of the mechanism of action of PX-478. B Western blot analysis of HIF-1α, BNIP3, NIX, DRP1, LC3B, and p62 protein levels in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. C Autophagic flux was quantified by colocalizing LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP levels were determined in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. G CCK-8 assay showing cell viability in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. H Cell apoptosis was measured in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia using Annexin V-FITC staining. I Western blot analysis of C-Caspase-3 and BAX protein levels in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia using an ELISA kit. M9a+sh EPAS1 cells ( n = 4) were treated with 30 μM PX-478 (for the last 22 h) under hypoxia (1% O₂) for 48 h. #1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. Two-tailed, paired Student’s t tests; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Schematic diagram of the mechanism of action of Mdivi-1. B Western blot analysis of DRP1, NIX, BNIP3, LC3B, and p62 protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. C Autophagic flux was assessed by quantifying the colocalization of LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP content were measured in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. G CCK-8 assay showing cell viability in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. H Cell apoptosis was determined in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia using Annexin V-FITC staining. I Western blot analysis of C-Caspase-3 and BAX protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) using immunofluorescence in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured using ELISA kits in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. M9a+sh EPAS1 cells ( n = 4) were treated with 50 μM Mdivi-1 or DMSO (pre-treatment for 4 h, followed by treatment for 48 h) under hypoxia (1% O₂).#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. Two-tailed, paired Student’s t tests; P < 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test